Progression of DCIS to invasive breast cancer through CCR2 chemokine signaling

通过 CCR2 趋化因子信号传导将 DCIS 进展为浸润性乳腺癌

• 批准号: 9093752
• 负责人: FARIBA BEHBOD
• 金额: $ 41.23万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9093752
• 关键词: Affect; Apoptotic; Biology; Breast; Breast Cancer Patient; Breast Cancer cell line; Breast Carcinoma; CC chemokine receptor 2; CCL2 gene; Carcinoma; Cell Culture System; Cell Line; Cell Survival; Cells; Development; Disease Progression; Ductal; Ductal Carcinoma; Fibroblasts; Goals; Health; Human; In Situ Lesion; Injection of therapeutic agent; Lead; Lesion; Ligand Binding; MAP Kinase Gene; Malignant Epithelial Cell; Mammary Neoplasms; Mammary gland; Mediating; Modeling; Molecular; Mus; Non-Invasive Lesion; Noninfiltrating Intraductal Carcinoma; Normal tissue morphology; Pathway interactions; Patients; Phenotype; Proteins; Receptor Signaling; Risk; Role; Sampling; Signal Transduction; Staging; Study models; Therapeutic; Transplantation; Woman; Xenograft Model; autocrine; base; biochemical model; breast cancer diagnosis; capsule; cell motility; chemokine; chemokine receptor; ductal breast carcinoma; in vivo; infiltrating duct carcinoma; insight; knock-down; macrophage; malignant breast neoplasm; mouse model; novel; outcome forecast; overexpression; paracrine; personalized approach; pro-apoptotic protein; protein expression; three dimensional cell culture; tumor; tumor progression

DESCRIPTION (provided by applicant): Ductal carcinoma in situ (DCIS) is the most common type of non-invasive breast cancer diagnosed in women and an immediate precursor to invasive ductal carcinoma (IDC). Current cyto- and histopathological approaches do not accurately predict disease progression, resulting in patients being under-treated or over-treated for DCIS. Our long-term goals are to identify key factors that lead to IDC that will enable the development of a molecular based approach to predict the risk of IDC, and a more tailored approach to treat DCIS. CCL2/CCR2 chemokine receptor signaling in DCIS is a potentially important mechanism for DCIS progression to IDC. CCR2 and its binding ligand CCL2, are overexpressed in breast ductal carcinomas, and correlate with tumor grade and poor patient prognosis. CCL2/CCR2 chemokine signaling is best known for regulating macrophage recruitment. However, knockdown of CCL2 significantly inhibits mammary tumor progression without significantly affecting macrophage recruitment. Overexpression of CCR2 in ductal carcinoma cells enhances cell survival and invasiveness, associated with increased expression of pro-survival and pro-invasive proteins (ALDH1A1) and decreased expression of pro-apoptotic proteins (Htra2). These studies indicate an important role for CCR2 signaling in breast ductal carcinoma progression. To clarify the in vivo role of CCL2/CCR2 signaling, we have developed novel xenograft models through mouse Mammary intraductal injection (MIND) of human breast carcinoma cell lines, Sum225 and DCIS.com. The Sum225 MIND model form stable non-invasive lesions, while DCIS lesions progress to IDC in the DCIS.com MIND model. Compared to conventional transplant models, these MIND models more closely mimic the biology of DCIS found in patients. Compared to Sum225 lesions, DCIS.com lesions show higher expression of CCL2, CCR2 and ALDH1A1 proteins and lower expression of Htra2, correlating with progression to invasive carcinoma. Based on preliminary studies, we hypothesize that CCL2/CCR2 signaling in ductal carcinoma cells enhances progression of DCIS to IDC. Aim 1: To characterize the role of autocrine CCL2/CCR2 signaling in ductal carcinoma cells during DCIS by analyzing effects of CCR2 overexpression and knockdown in MIND models, 3D cell culture models and biochemical approaches, using primary and established breast cancer cell lines. Aim 2: To determine the functional contribution of stromal derived CCL2 on CCR2 mediated progression of DCIS, by analyzing the effects of CCL2 derived from fibroblasts on DCIS progression, using subrenal graft transplant models, MIND models, 3D cell culture models and biochemical approaches, using primary and established breast cancer cell lines. Through a multiple PI plan, the objective is to determine how CCL2/CCR2 signaling regulates expression of pro-invasive and pro-survival factors (ALDH1A1) and pro-apoptotic factors (Htra2) to promote ductal carcinoma progression. These studies will reveal mechanistic insight into the role of chemokine receptor signaling in DCIS, with important therapeutic implications. .

描述（由申请人提供）：导管原位癌（DCIS）是女性中最常见的一种非浸润性乳腺癌，是浸润性导管癌（IDC）的直接前兆。目前的细胞和组织病理学方法不能准确预测疾病进展，导致DCIS患者治疗不足或过度治疗。我们的长期目标是确定导致IDC的关键因素，从而开发基于分子的方法来预测IDC的风险，以及更有针对性的治疗DCIS的方法。DCIS中CCL2/CCR2趋化因子受体信号传导是DCIS发展为IDC的潜在重要机制。CCR2及其结合体CCL2在乳腺导管癌中过表达，并与肿瘤分级和患者预后不良相关。CCL2/CCR2趋化因子信号最广为人知的是调节巨噬细胞募集。然而，敲低CCL2可显著抑制乳腺肿瘤的进展，而不显著影响巨噬细胞的募集。CCR2在导管癌细胞中的过表达可提高细胞存活率和侵袭性，与促生存和促侵袭蛋白（ALDH1A1）的表达增加和促凋亡蛋白（Htra2）的表达减少有关。这些研究表明CCR2信号在乳腺导管癌进展中的重要作用。为了阐明CCL2/CCR2信号在体内的作用，我们通过小鼠乳腺导管内注射（MIND）建立了人类乳腺癌细胞系Sum225和DCIS.com的新型异种移植模型。Sum225 MIND模型形成稳定的非侵袭性病变，而DCIS在DCIS.com MIND模型中进展为IDC。与传统的移植模型相比，这些MIND模型更接近于模拟患者DCIS的生物学特性。与Sum225病变相比，DCIS.com病变CCL2、CCR2和ALDH1A1蛋白表达较高，Htra2表达较低，与浸润性癌的进展相关。基于初步研究，我们假设导管癌细胞中的CCL2/CCR2信号通路促进DCIS向IDC的进展。目的1：通过在MIND模型、3D细胞培养模型和生化方法中分析CCR2过表达和敲低对原发性和已建立的乳腺癌细胞系的影响，表征自分泌CCL2/CCR2信号在DCIS期间导管癌细胞中的作用。目的2：利用原代和已建立的乳腺癌细胞系，采用肾下移植模型、MIND模型、3D细胞培养模型和生化方法，分析成纤维细胞来源的CCL2对DCIS进展的影响，以确定基质来源的CCL2对CCR2介导的DCIS进展的功能贡献。通过多次PI计划，目的是确定CCL2/CCR2信号如何调节促侵袭和促生存因子（ALDH1A1）和促凋亡因子（Htra2）的表达，促进导管癌的进展。这些研究将揭示趋化因子受体信号在DCIS中作用的机制，具有重要的治疗意义。